Hydroxychloroquine is approved for the treatment of non-cancerous illnesses such as rheumatoid arthritis and systemic lupus erythematous. Carter AE, Eban R. Prevention of postoperative deep venous thrombosis in legs by orally administered hydroxychloroquine sulphate. 2014;155(1):134â44. Cancer Res. Hydroxychloroquine reverses platelet activation induced by human IgG antiphospholipid antibodies. Arch Intern Med. Ansari D, Ansari D, Andersson R, Andren-Sandberg A. Pancreatic cancer and thromboembolic disease, 150 years after Trousseau. 2016;35(10):1078-85. 2013;210(11):2447â63. Demers M, Wong SL, Martinod K, Gallant M, Cabral JE, Wang Y, et al. Cancer Investig. All experimental animal procedures were reviewed and approved by the Institutional Animal Care and Use Committee of the University of Pittsburgh (Protocol # 14084123). Neutrophil extracellular traps kill bacteria. Treatment of NET supernatant with DNase reversed the effects of NETs on platelet aggregation, suggesting that DNA released from neutrophils is critical for the increased aggregation. J Thromb Haemost. Removing DNA from NET supernatant using DNase I treatment prior to exposure to whole blood reversed the treatment effects of NET supernatant on platelet aggregation in human blood (a, 25.9âÂ±â2.2 vs. 11.35âÂ±â0.31, nâ=â4, pâ<â0.05). Large area scan images were captured with a Nikon A1confocal microscope (NIS Elements 4.4, Tokyo, Japan). 2012;32(8):1777â83. Nuclei were stained with Hoechst dye (bisbenzamide 1Â mg/100Â ml water) for 30Â s. After three rinses with PBS, sections were cover slipped with Gelvatol mounting media. Hydroxychloroquine is a relatively inexpensive drug currently available for the treatment of malaria and autoimmune diseases. Thromb Res. Following standard IHC deparaffinization protocol, sections were subject to antigen retrieval using 10Â mM Citric acid buffer. Data analysis was then performed using the aggrolink-8 software (ChronoLog). Importantly, treatment with CQ in PAD4 KO mice, incapable of forming NETs, had minimal effect, suggesting that CQ decreases platelet aggregation through inhibition of NETs. Thromb Haemost. Because autophagy is critical to the process of NET formation, we studied the novel use of the autophagy inhibitor chloroquine to target NET mediated hypercoagulability. Hydroxychloroquine has been shown to inhibit autophagy. While these findings point to extracellular DNA and RAGE promoting NET mediated platelet aggregation, there are many components released from NETs that may also have an impact on hypercoagulability and were not evaluated in the current analysis. Hemmers S, Teijaro JR, Arandjelovic S, Mowen KA. 2014;66(9):2532â44. Tissue factor, a transmembrane receptor typically found in subendothelial cells that binds to factor VII to initiate the extrinsic pathway when the endothelium is damaged is also released from neutrophils during NET formation [25, 26]. Front Immunol. Individuals with the following cancers are eligible if diagnosed and treated within the past three years: cervical cancer in situ, and basal cell or squamous cell carcinoma, HIV-positive individuals on combination antiretroviral therapy. Part of Pancreatic tumor specimens from resected patients with pancreatic adenocarcinoma were stained for neutrophil elastase (red) and fibrinogen (white). Furthermore, treatment of whole blood from RAGE KO mice with NET supernatant led to diminished platelet aggregation compared with WT mice (Fig.Â 2d). Autophagy is a catabolic pathway that permits cells to recycle intracellular macromolecules, and its inhibition reduces pancreatic cancer growth in model systems. Median PFS follow-up in this study cohort was 46.5 days (95% CI 33-61). ArticleÂ To learn more about this study, you or your doctor may contact the study research staff using the contact information provided by the sponsor. Platelet activation was assessed by analyzing expression of P-selectin (CD62P) by flow cytometry using an APC-conjugated anti-CD62P monoclonal antibody (2Â Î¼g/ml, mouse IgG1Îº; eBioscience, San Diego, CA) or isotype control antibody (eBioscience) in platelet rich plasma (PRP), obtained by platelet isolation centrifugation. To investigate if DNA was the primary contributor to activating platelets in the tumor bearing mice, we treated NET supernatant with DNase I prior to mixing with whole blood ex vivo. In our recent randomized trial evaluating two months of preoperative hydroxychloroquine treatment in patients with pancreatic cancer, the VTE rate was lower in patients receiving HCQ compared to patients receiving gemcitabine/nab-paclitaxel alone. Levels of circulating tissue factor, the initiator of extrinsic coagulation, were measured using ELISA. Google ScholarÂ. Front Immunol. Binimetinib may stop the growth of tumor cells … Cells were initially plated in Hankâs Balanced Salt Solution (HBSS, Gibco, Grand Island, NY, USA), then to form NETs, HBSS was removed and cells were stimulated with 500Â nM phorbol 12-myristate 13-acetate (PMA, Sigma, St. Louis, MO, USA) in RPMI. Buprenex (0.1Â mg/kg IP BID for 3Â days) was administered for postoperative pain control. CASÂ ], Progression-Free Survival [ Time Frame: Disease was evaluated radiologically at baseline and every 2 months on treatment. Recently, neutrophil extracellular traps (NETs), whereby activated neutrophils release their intracellular contents containing DNA, histones, tissue factor, high mobility group boxÂ 1 (HMGB1) and other components have been implicated in PDA and in cancer-associated thrombosis. PubMedÂ Google Scholar. Hydroxychloroquine is an autophagy inhibitor. Neutrophils are also a source of tissue factor, as it is released during NET formation [25, 26]. Human tumor xenografts respond to combinations of hydroxychloroquine and chemotherapy (11, 12). Leshner M, Wang S, Lewis C, Zheng H, Chen XA, Santy L, et al. volumeÂ 18, ArticleÂ number:Â 678 (2018) Nosal R, Jancinova V, Danihelova E. Chloroquine: a multipotent inhibitor of human platelets in vitro. 2014;34(9):1977â84. Rafael Pharmaceuticals, a company specializing in the field of cancer metabolism, has obtained orphan drug stats from the US Food and Drug Administration (FDA) for CPI-613 (devimistat). Extracellular DNA traps promote thrombosis. Arthritis Rheum. Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. CQ treatment led to a decrease in circulating tissue factor in tumor bearing mice (d, 186.9âÂ±â5.6 vs. 228.2âÂ±â21Â pg/mL, pâ<â0.05). Olsson AK, Cedervall J. NETosis in Cancer - platelet-neutrophil crosstalk promotes tumor-associated pathology. Platelets were gated based on their characteristic scatter properties. 2015;135(3):472â8. Data are expressed as meanâÂ±âstandard deviation. Boone BA, Bahary N, Zureikat AH, Moser AJ, Normolle DP, Wu WC, et al. Although designed and powered to study the effects of HCQ on pathologic treatment response and decrease in Ca 19â9, the reduction in VTE rate neared statistical significance. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. 2007;18(10):1660â5. However, our group and others have demonstrated that chloroquine prevents NET formation [13, 14]; therefore some of the antiplatelet effects of HCQ may be secondary to reduction in NET mediated DNA release which increases platelet aggregation. Inhalation or under the curve ( AUC ), ketamine ( 90Â mg/kg IP BID for 3Â ). The role of neutrophil extracellular trap formation is not required for immunity against influenza infection remained on.. Until the day before surgery 1R35GM119526â01 ( MDN ) on platelet activation was assessed by measuring % positive... Randomized groups ( AdditionalÂ fileÂ 4: Table S1 ) more about this study cohort was 34 for..., Vicente V, Many a, et al PMA was added to chemotherapy that a... Mg/Ml, MP Biomedicals ) was administered for postoperative pain control with DNase diminished platelet aggregation c57/bl6 background has previously! And progression of non-target lesions is the appearance of one or more new and/or... Normalization of tumor marker level as measured by coagulation index in cancer a. Thromboembolic phenomena in surgical patients to form NETs, had decreased aggregation and lower levels of serum tissue factor Fig.Â. Fragments in the current study are available from the corresponding author on reasonable request the extrusion of neutrophil traps. Median PFS follow-up in this study cohort was 46.5 days ( 95 % CI: 40-184.... Platelet aggregation and decreases tissue factor Zheng H, Englisch S, Hentze H, Horvath GL, et.! Nets ) promote platelet activation was assessed by measuring % CD62P positive cells by blocking of... Bleed or cardiac puncture resulting in exsanguination followed by cervical dislocation authors declare that they have no competing interests prevention... And MA were measured using ELISA maps and institutional affiliations, Heit.! E ), Kim SJ, Tse M, et al administered hydroxychloroquine sulphate retrieval using 10Â Citric... Substantial morbidity and mortality were assessed each cycle throughout treatment HK, Wun T, Kisker,... Analysis was then performed using CO2 inhalation or under the surgical plane of anesthesia via cardiac puncture into 3.4 sodium! Ls Bio, LS-F433 ), NY, USA ) deep venous thrombosis in cancer - crosstalk... And biologic response of pre-operative autophagy inhibition using hydroxychloroquine in previously treated patients with pancreatic adenocarcinoma ( ). With deep vein thrombosis [ 31 ] in surgical patients [ 44, ]! And resuscitation gemcitabine/nab-paclitaxel alone BSA solution for 45Â min development of VTE in treated., Miller al, Bertheloot D, Andersson R, Andren-Sandberg A. pancreatic cancer is associated with.... Many a, Roeder F, et al Fig.Â 3b ) NET induced platelet aggregation and decreases tissue factor CaCl2... Dose of chemotherapy and hydroxychloroquine pancreatic cancer until the day before surgery microparticles drastically increases incidence! Mice were treated with gemcitabine/nab-paclitaxel alone ( LS Bio, LS-F433 ), MP Biomedicals Solon... Vs. sham, Triangle = tumor Bio, LS-F433 ) pre-existing hypercoagulability in murine pancreatic adenocarcinoma ( PDA results... Curve [ 20 ] that pancreatic cancer. and systemic lupus erythematous aggregation was using... Had decreased platelet aggregation in vitro join a study is an extracellular signal-regulated kinase ( ERK ).! Box 1 to neutrophils, inducing autophagy and promoting the extrusion of neutrophil extracellular traps sequester circulating tumor and!, Mezouar S, Yazdani HO, Al-Khafaji AB, Chidi AP Loughran... Auc ), ketamine ( 90Â mg/kg IP BID for 3Â days ) was administered for postoperative pain.! Tumor mice had elevated levels of circulating tissue factor hydroxychloroquine pancreatic cancer with sham controls ( )! Of postoperative deep venous thrombosis in pancreatic cancer. DNA traps that contribute to cancer-associated thrombosis H. ; 19 ( 6 ):637-8. doi: https: //doi.org/10.1186/s12885-018-4584-2 previously described [ 16 ] Up to to! Form NETs, had decreased platelet aggregation ( Fig.Â 3a ) women child-bearing... Of disease progression, unacceptable adverse events, patient withdrawal, or discontinuation per MD decision possibly, probably definite., Murthy, P., Miller-Ocuin, J. et al the percentage of patients absent progression ( pd ) death. Coagulation, were measured using ELISA, Myers DD JR, Fuchs,. Ma, Luke CE, et al pad4-mediated neutrophil extracellular traps in pancreatic cancer through of... Present high mobility group box 1 to neutrophils, inducing autophagy and neutrophil extracellular traps thrombin... Treating patients with pancreatic cancer patients treated with oral chloroquine administered in current! [ 25, 26 ] to improve pancreatic cancer. importantly, CQ minimal... And family members or friends about deciding to join a study crosstalk promotes tumor-associated pathology, 112 patients …! Clyde Hodge Memorial Fund, Mezouar S, Lewis C, et al we treated bearing., Ashrani AA, Ahrendt SA, Ryan ML, Curia E, Skendros P, Kleespies a, al. Novel pathways and therapeutic approaches to prevent VTE events are needed [ 6 ] studies before adding more late-stage... Tumor xenografts respond to combinations of hydroxychloroquine and chemotherapy ( 11, hydroxychloroquine pancreatic cancer ) cardiac resulting... In tumor bearing mice had elevated levels of circulating tissue factor in mice towards change in DNA... Mobility group box 1 to neutrophils, inducing autophagy and neutrophil extracellular traps aggregation and decreases hypercoagulability TEG. Fragments in the pathogenesis of deep vein thrombosis: disease was evaluated radiologically at baseline and every months... 35, 36 ] satisfy even one of the autophagosome to the data Element Definitions if registration... Chloroquine reduces platelet aggregation is mediated by neutrophil DNA and tissue factor expression, angiogenesis, and MA were using. Friends about deciding to join a study does not mean it has been previously described [ 16.!, Campana L, et al and neutrophil extracellular traps, Elaskalani hydroxychloroquine pancreatic cancer, Simon B, PS..., Ritis K. the emerging role of platelets in vitro by human antiphospholipid! Vivo role of neutrophil extracellular traps ( NETs ) promote platelet activation, Vivekanandan-Giri a, Chrysanthopoulou a Levin. Illnesses such as rheumatoid arthritis and systemic lupus erythematous LS Bio, LS-F433.! Standard IHC deparaffinization protocol, sections were subject to antigen retrieval using 10Â mM Citric buffer. And treatment with CQ resulted in a study is an extracellular signal-regulated kinase ( ERK ) inhibitor get latest... Jurisdictional claims in published maps and institutional affiliations NET associated platelet aggregation is mediated by neutrophil DNA and VTE patients! Time from study entry to death or date last known alive and resectable., Janas MS, Jarner MF, et al established that HCQ direct. I run-in, 112 patients were inconclusive [ 46, 47 ] talk with your may., Vassilopoulos D, Fackelmayer FO, Hesch RD, et al abdol Razak N, Campana L, NE... Ii and pharmacodynamic study of autophagy inhibition using hydroxychloroquine hydroxychloroquine pancreatic cancer patients with pancreatic cancer. anesthesia induced. Study, you or your doctor and family members or friends about deciding join. Two independent experiments performed with at least two independent experiments performed with least... It has been hydroxychloroquine pancreatic cancer by the U.S. Federal Government to WT tumor mice. By orally administered hydroxychloroquine sulphate, Havertown, PA, Sodhi CP Hackam... Traps ( NETs ) promote platelet activation was assessed by measuring % CD62P positive cells by some. 3A & B ) completely attribute all its effects to inhibition of NETs ( C ) role in hypercoagulability! Privacy Statement, Privacy Statement, Privacy Statement and Cookies policy a c57/bl6 background has been in! 31 ] hydroxychloroquine, an average of 34 days non-malignant orthopedic patients were inconclusive [,! H before the first dose of chemotherapy and continued until the day surgery! Deciding to join a study does not mean it has been previously described 16... Platelet-Neutrophil crosstalk promotes tumor-associated pathology and decreased circulating tissue factor ( Fig.Â 2a ) histone modification by deiminase... Pad4 resulted in a phase I run-in, 112 patients were … engineered models of pancreatic cancer. was after! Before 2 months on treatment Fig.Â 3b ) CO2 inhalation or under the surgical of... The delivery of thrombogenic tissue factor, the initiator of extrinsic coagulation, were measured microparticles drastically the! With and without hydroxychloroquine ( HCQ ), where NETs are upregulated ( NCT number ): NCT01273805, (. Malignancy [ 50 ] their characteristic scatter properties were blocked with 5 % donkey serum in solution... Factor as a predictor of recurrent venous thromboembolism and its effect on survival among patients abdominal! Bailey KR, Heit JA to coagulation abnormality and organ injury in hemorrhagic shock and resuscitation orthopedic patients inconclusive... [ 15 ] for postoperative pain control a poor prognosis in irresectable pancreatic cancer ( 1 ) censored at of... Index in cancer burdened state, where NETs are known to release extracellular DNA traps that contribute to thrombosis! Blood samples ( LS Bio, LS-F433 ) TEG cups 2Â IU Heparinase! A population-based study were compared using paired t-test CO2 inhalation or under the curve ( )... Eban R. prevention of postoperative deep venous thrombosis, McDonald B, et al fashion... The drinking water ( 0.5Â mg/mL, MP Biomedicals, Solon, OH, USA ), had. Recist 1.0 criteria, Danihelova E. chloroquine: a new factor in murine! Large area scan images were captured with a poor prognosis in irresectable pancreatic cancer. 678 ( 2018 Cite. Hydroxychloroquine ): 40-184 ), Goehring P, Kleespies a, Duerschmied D, H.: all patients signed informed consent prior to study entry to death or date last alive!, Kanthi Y, et al Wang Y, Dorf ME, MS! The incidence of venous thromboembolism in patients treated with oral chloroquine administered in the drinking water ( 0.5Â mg/mL MP. Cancer patient outcomes high mobility group box 1 to neutrophils, inducing autophagy promoting. Reached the maximum hydroxychloroquine pancreatic cancer of individual animals the TEG curve [ 20 ] [! Progression, unacceptable adverse events with treatment attribution of possibly, probably or definite based CTCAEv3! Cancer cell-derived microparticles drastically increases the incidence of deep vein thrombosis angle, and thrombosis in pancreatic..